title: Toxicity assessments of nonsteroidal anti-inflammatory drugs in isolated mitochondria, rat hepatocytes, and zebrafish show good concordance across chemical classes.
abstract: To reduce costly late-stage compound attrition, there has been an increased focus on assessing compounds in in vitro assays that predict attributes of human safety liabilities, before preclinical in vivo studies are done. Relevant questions when choosing a panel of assays for predicting toxicity are (a) whether there is general concordance in the data among the assays, and (b) whether, in a retrospective analysis, the rank order of toxicity of compounds in the assays correlates with the known safety profile of the drugs in humans. The aim of our study was to answer these questions using nonsteroidal anti-inflammatory drugs (NSAIDs) as a test set since NSAIDs are generally associated with gastrointestinal injury, hepatotoxicity, and/or cardiovascular risk, with mitochondrial impairment and endoplasmic reticulum stress being possible contributing factors. Eleven NSAIDs, flufenamic acid, tolfenamic acid, mefenamic acid, diclofenac, meloxicam, sudoxicam, piroxicam, diflunisal, acetylsalicylic acid, nimesulide, and sulindac (and its two metabolites, sulindac sulfide and sulindac sulfone), were tested for their effects on (a) the respiration of rat liver mitochondria, (b) a panel of mechanistic endpoints in rat hepatocytes, and (c) the viability and organ morphology of zebrafish. We show good concordance for distinguishing among/between NSAID chemical classes in the observations among the three approaches. Furthermore, the assays were complementary and able to correctly identify "toxic" and "non-toxic" drugs in accordance with their human safety profile, with emphasis on hepatic and gastrointestinal safety. We recommend implementing our multi-assay approach in the drug discovery process to reduce compound attrition.
authors.affiliation: Compound Safety Prediction, Worldwide Medicinal Chemistry, Pfizer, Inc., Groton, CT 06340, USA.
authors.forename: Sashi
authors.initials: S
authors.lastname: Nadanaciva
authors.forename: Michael D
authors.initials: MD
authors.lastname: Aleo
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Strock
authors.forename: Donald B
authors.initials: DB
authors.lastname: Stedman
authors.forename: Huijun
authors.initials: H
authors.lastname: Wang
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Will
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000737:chemistry* / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000473:pathology; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D019055:No-Observed-Adverse-Effect Level; D010101:Oxygen Consumption / Q000187:drug effects; D061251:Primary Cell Culture; D051381:Rats; D018675:Toxicity Tests / Q000379:methods*; D015027:Zebrafish / Q000196:embryology
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal
keywords: AC(50); AERS; COX; ER; GI; High content imaging; IC(50); LOEC; Liver toxicity; Mitochondrial toxicity; NOEC; NSAID; NSAIDS (non-steroidal anti-inflammatories); ROR; UC(50); Zebrafish; adverse event reporting system; cyclooxygenase; endoplasmic reticulum; gastrointestinal; half-maximal activity concentration; lowest observed effect concentration; no observed effect concentration; nonsteroidal anti-inflammatory drug; reporting odds ratio; the concentration at which 50% inhibition occurred in State 3 respiration; the concentration at which 50% uncoupling occurred in State 2 respiration